FIGURE 3.

FAM60A promotes the retention of HDAC1 but not SIN3 at the cyclin D1 promoter in asynchronously growing cells. A and B, overexpression of SIN3A represses cyclin D1. U2OS cells stably expressing GFP-tagged SIN3A in a tetracycline-inducible manner were transfected with two separate FAM60A siRNAs or with a control (GFP-specific) siRNA. Cells were incubated, or not, with tetracycline. Cyclin D1 mRNA levels were quantitated by quantitative PCR (A), and cell extracts were subjected to Western blotting with the indicated antibodies (B). Data are represented as means ± S.E. (error bars) of triplicate experiments. C, extracts of cells from A were subjected to ChIP analysis with the indicated antibodies. Quantitative PCR was carried out on DNA extracted from the precipitates (ip) with primers specific for the cyclin D1 promoter (left panels) or the GAPDH promoter (right panels). D, extracts of cells from A were subjected to ChIP analysis with the indicated antibodies. Quantitative PCR was carried out on DNA extracted from the precipitates with primers specific for the ALDH6A promoter.